Compositions having testosterone antagonistic effects and method of using the same

ABSTRACT

A NEW USE FOR KNOWN COMPOUNDS OF THE CATEGORY 2LOWER ALKANOYLAMINO-5-NITROTHIAZOLES IS DESCRIBED. THESE COMPOUNDS, WHEN ADMINISTERED TO MAMMALS, COUNTERACT THE BIOLOGICAL EFFECT OF TESTOSTERONE ON END ORGANS.

United States Patent 3,644,628 COMPOSITIONS HAVING TESTOSTERONE AN- TAGONISTIC EFFECTS AND METHOD OF USING THE SAME George Tonelli, 15 Seneca Ave., Emerson, NJ. 07630 No Drawing. Continuation-impart of application Ser. No.

736,982, June 14, 1968, which is a continuation-inpart of application Ser. No. 628,295, Apr. 4, 1967. This application Aug. 29, 1969, Ser. No. 854,284

Int. Cl. A61v 27/00 U.S. Cl. 424--270 6 Claims ABSTRACT OF THE DISCLOSURE A new use for known compounds of the category 2- lower alkanoylamino-S-nitrothiazoles is described. These compounds, when administered to mammals, counteract the biological eifect of testosterone on end organs.

This application is a continuation-in-part of my application Ser. No. 736,982, filed June 14, 1968, which in turn is a continuation-in-part of my application Ser. No. 628,295, filed Apr. 4, 1967, both are now abandoned.

BRIEF SUMMARY OF THE INVENTION This invention relates to a method of administering pharmaceutical compositions. More particularly, to administering novel pharmaceutical compositions containing lower alkanoyl aminonitriothoiazoles which are eflective in decreasing the biological effect of testosterone on end organs (anti-testosterone effect). The said compositions are of potential use in the therapy of endocrinopathies related to excess endogenous production or end organ hyper-responsiveness to testosterone.

The present invention comprises a pharmaceutical composition containing as the essential active component, a compound of the formula:

N-CH wherein R is lower alkanoyl. Among the compounds included in the scope of the invention are, for example, 2- acetylamino-5-nitrothiazole, 2-propionylamino 5 nitrothiazole or Z-butyrylamino-5-nitrothiazole.

The 2-lower alkanoylamino 5-nitrothiazoles of the present invention are, in general, greenish-yellow to orange-yellow to yellow colored crystalline solids, relatively insoluble in water.

The compounds are well known in the literature but up until the time of this invention, their use for antitestosterone effect has not been described. Methods for the preparation of these compounds are found in the literature, such as US. Pat. 2,531,756 or 2,631,963. The active components of the compositions of matter of the present invention, the 2-loweralkanoylamino-S-nitrothiazoles, have been found to be highly efiective in restricting the biological efiect of testosterone in mammals, for example, rats and mice. The doses at which these effects occurred, ranged from about 1 to 64 mg./kg./day, which are substantially lower than effective antitrichomoniasis doses. In this connection, as noted in the recent review Advances in Chemotherapy, vol. 3, 1968, Academic Press, New York and London, Eds. A. Goldin, F. Hawking and R. J. Schnitzer, the curative dose in the rat for Trichomonas 3,644,628 Patented Feb. 22, 1972 vaginalis was greater than 300 mg./kg./day by the oral route. Curative doses of between 300-400 mg./ kg. against this same organism are described in Experimental Chemotherapy vol. 1, 1963. Eds. Schnitzer, R. 1., and Hawking, F., Academic Press, New York.

The compounds do not present any lasting or serious side effects and have satisfactory ranges between therapeutically elfective and toxic doses in mammals.

The findings (see Examples 1, 2, 3 and 4) that these compounds are capable of reducing the hypertrophy of certain end organs resultant from administration of testosterone is of practical importance in the treatment of syndromes caused by excess testosterone production.

The compositions of the present invention containing the active component and a non-toxic carrier may be administered orally or parenterally as well as topically. For oral or parenteral administration the compositions are preferably presented for administration in unit dosage forms, such as, for example, tablets, capsules, granules, powders, oral solutions or suspensions, or sterile parenteral solutions or suspensions, and the like. The amount of active component administered per day may vary from 1 to 64 mg./ kg. of mammal.

For preparing solid compositions such as tablets, the active component is mixed with conventional tableting ingredients such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate, gums and functionally similar materials as pharmaceutical diluents or carriers. The tablets of the novel compositions can be laminated or otherwise compounded to provide a dosage form aifording the advantage of prolonged or delayed action or predetermined successive action of the enclosed medication. For example, the tablet can comprise an inner dosage and an outer component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer which serves to resist distintegration in the stomach and permits the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such as materials including a number of polymeric acids or mixtures of polymeric acids with such materials as shellac, shellac and cetyl alcohol, cellulose acetate and the like. A particularly advantageous enteric coating comprises a styrene maleic acid copolymer together with known materials contributing to the enteric properties of the coating.

The liquid forms in which the novel compositions of the present invention may be incorporated for administration include aqueous solutions, suitably flavored syrups, aqueous or oil suspensions, flavored emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil, peanut oil, and the like, as well as elixirs and similar pharmaceutical vehicles. Suitable dispersing or suspending agents for aqueous suspensions include synthetic and natural gums such as tragacanth, acacia, alginate, dextran, sodium carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone, gelatin, and the like. Sterile suspensions or solutions are required for parenteral use. Isotonic preparations containing suitable preservatives are also highly desirable for injection use.

The term unit dosage form as used in the specification and claims refers to physically discrete units suitable as unitary dosages for mammals each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic elfect in association with the required pharmaceutical diluent, carrier or vehicle. A convenient dosage unit form may contain 25 to 250 milligrams of the active therapeutic ingredient.

3 DETAILED DESCRIPTION The following examples describing specific testing proly at doses ranging from 2 to 30 mg./kg. per rat. The compound exhibits significant anti-testosterone activity.

TABLE I Anti-testosterone activity of test compound as assessed in the castrated rat receiving 50 or 100 mcg. of testosterone Relative organ wts. (mg/100 g.)

Percent Percent Dose of Rat weight (g.) testostestosantl-androgen, terone terone Treatment megJrat/day Initial Final Vent. Prost. response Sern. Vesc. response CMC vehicle 63:1:7 925:10 10. 2&4 11. :i:2 6317 100:1:10 43. 8:1:11 100 56. 9:1:16 100 Testosterone (50 meg) 100 635:6 9718 31. 85:; g 22 32;.19gt1g 3 egg 2- t l itrothia ole- 500 633:5 935:9 21. 8d: :1: Testosterone (50 meg) plus ace ylam no n z I 500 63i5 94:7 23. 3:129 a 53 22. 9&5 a 40 CMC vehicle 65:1:4 9918 9. 45:2 10. 3:1:2 66i4 1025:!) 67. 4:]:13 100 90. 9:1:23 100 Testosterone (100 mcg 672E; 1052a 49' his i g t the ole 500 67:1:4 105:l:6 52. 5:|: 0 Testosterone (100 meg) plus 2 aeetylamino 5 m 1 z 2, 500 6H2 983:9 43. 85:5 3 65 44' E213 a 49 NoTE.Vent. Prost.=ventral prostate. Sem. Vese.=seminal vesicles. Testosterone and test compounds administered subcutaneously 4 days after castration once a day, for 7 consecutive days. Values given are means i standard deviation. Eight rats used as controls and with each dosage level.

EXAMPLE 2 Table II shows that 2-acetylamino-S-nitrothiazole reduces the eifects of testosterone in the castrated mouse as well. The use of the castrated mouse is likewise noted in the literature reference given under Example 1. In this bioassay, the subject compound is administered orally once a day for 7 consecutive days. Testosterone is administered concomitantly at daily doses of 100 mcg. by subcutaneous route. The compound exhibits significant anti-testosterone action.

TABLE 11 Effects of Q-acetylamino-5-nitrothiazole administered subcutaneously with and without testosterone, to the hemicastrated rat for ten consecutive days Rat wt. (g). Relative organ weights (mg/100 g.) Dose, No. of Treatment mglratlday rats Initial Final Testis Vent. Prost. Sem. Vese.

CMC (intact) 8 57. 1 102. 2 473. 5:};57 6. 7:1:7 20. 8:1:7 CMO 8 55. 3 101. 2 530. 9:1:50 45. 9i13 18. 7:|:6 8 55. 6 91. 1 543. 0&85 43. 0i7 13. 7i4 2-acetylamino-5-nitrothiazole 8 57. 5 98. 3 551. 0:l:82 43. 9:|:7 15. 53:4 8 65. 6 106. 0 508. :43 39. 2:i:7 14. 5:l:6 Testosterone (100 meg.) 8 59. 6 106. 0 288. 65:49 101. 7i23 118. 05:29 6 59. 2 96. 1 514. 2166 2 57. :14 2 28. 8i4 Testosterone (100 mcg.) plus 2-acetylamino-5-nitrothiazole 6 59. 0 87. 6 455. 4i69 2 71. 8:1:10 Z 28. 65:8 6 57. 8 84. 0 386. li88 2 55. 35:14 2 31. 2d: 7

No'rn.-Vent. Prost.=ventra1 prostate. Sem. Vese.=seminal vesicles. Values given are means i standard deviation.

examples amounts are by weight unless otherwise indicated.

EXAMPLE 1 Utilization of the castrated rat or mouse for assessing the potential anti-testosterone anti-androgen) effect of compounds is a recommended and accepted laboratory procedure, as noted in Methods in Hormone Research, vol. H, 1960, Academic Press, New York and London, Ed. -R. I. Dorfman, and in Experimental Endocrinology, 1964, Academic Press, New York and London, authors M. X. Zarrow, I. M. Yochim and I. L. McCarthy.

In this test, testosterone and the compound under examination are injected subcutaneously each at a separate site, once a day for 7 consecutive days, beginning on the fourth day following castration. Castration of the male removes the testes which are normally the major site of production and source of testosterone in the animal. The weights of seminal vesicles and ventral prostate, which depend on testosterone for their growth and integrity, are determined 24 hours after the last dosing. The compound is considered anti-testosterone if it interferes with this testosterone effect by reducing Weight gain, Table I summarizes the typical efiiects of 2-acetylamino-S-nitrothiazole administered subcutaneous EXAMPLE 3 Another method of determining the anti-testosterone activity of test compounds is with the use of the hemicastrated rat. This method, in addition, establishes whether the compound has antigonadotropic activity. Antigonadotropic activity is defined as inhibition of the release of the tropic hormones of the pituitary gland which controls testicular growth and whose action is independent, as we show, of the anti-testosterone effect of these claims. Four days after hemicastration (removal of right testicle), which results in greater gonadotropin release from the pituitary, rats receive testosterone and the test compound (at a separate site) once a day for 10 consecutive days. On the eleventh day, the remaining testicle, seminal vesicles and ventral prostate are removed and weighed. Testosterone alone causes an increase in the weights of the seminal vesicles and ventral prostate (androgenic effect), and a decrease in the weight of the remaining testicle (antigonadotropic effect). The results are shown in Table III. 2-acetylamino-5-nitrothiazole administered alone, did not reduce pituitary gonadotropin activity since testicular weight did not change; when administered together with testosterone, testicular weight was not further reduced as would be expected where the compounds are antigonadotropic.

Example 3 also shows Z-acetylamino-S-nitrothiazole inhibiting the weight increases of the ventral prostate and seminal vesicles which normally result from testosterone administration, again confirming the anti-testosterone activity of the compound.

TABLE III Anti-testosterone activity of test compound as assessed in the eastrated mouse receiving 100 mcg. of testosterone Norm-Vent. Prost =Ventral prostate. Sem. Vesc. Seminal vesicles. Values are means :1: standard deviation based on 7 animals per treatment EXAMPLE 4 A second method of determining if a compound has anti-gonadotropic as well as anti-testosterone effects is by the use of the parabiotic rat, as noted in standard textbooks of Endocrinology (references are given under Example 1). 1n the parabiosis study, 21-day old male Wyckotf rats, weighing an average of 45 g., are castrated and, 3 days later, surgically united to 23 day old intact female rats of an average weight to 50 g. (When united in this way the normal feed back effect on the pituitary of both the male and the female partners of endogenous testosterone produced by the testes is prevented and pituitary gonadotropins are released in a greater than normal amount, stimulating the growth of sensitively responsive tissues such as the ovaries.) The subject compound, either alone or with testosterone, is injected subcultaneously in the castrated male, once a day for 10 consecutive days. On the eleventh day the ovaries of the female partner and the ventral prostate and seminal vesicles of the male partner are removed and weighed. The data are presented in Table IV. Administration of 100 mcg./rat/day of testosterone reduced ovarian hypertrophy from 191:57 mg. to 64:29 mg., a reflection of the androgens known anti-gonadotropic activity. Ovarian weights following administration of 2 or 5 mg. of Z-acetylamino-S-nitrothiazole are not markedly changed. That 2- acetylamino-S-nitrothiazole does not possess anti-gonadotropic activity is substantiated by the results obtained following administration of the compound with testosterone. Under the latter conditions, 2-acetylamino-S-nitrothiazole significantly interferes with the ovarian weight depressing effects of testosterone. If the compound were anti-gonadotropic a further reduction in ovarian weight would have occurred. The data also show that 2-acetylamino-5- nitrothiazole reduces the effectiveness of testosterone on seminal vesicles and ventral prostate again confirming the anti-testosterone activities of the compound.

TABLE IV antigonadotropic and antlandrogenic activities of 2-acetylamino-5- nitrothiazole in the parabiotic rat Compounds administered Organ weighs: mg./100 g. B.W. (mg./rat/day) (Mean :1: S.D.)

2'acetyl- Castrated male amino-5- Intact Testosnitrothia- No. of female Seminal Ventral terone zoie pairs ovaries vesicles prostate 0 1 30:1:6 0 0 5 190i69 10. 8=i=2 12. 3:;2 0 2 5 159:1:73 10. 85:2 11. 4i4 0 0 1 13 32:1:5 0 0 13 191i57 12. 95:3 10. 6:1;5 0 5 12 175:|;49 12. 4:122 9. 712 0. 1 0 6 64=lz29 58. 05:10 54. 05:13 0. 1 2 4 2 l24:|;57 3 43. 0:1:14 Z 35. 0:1:7 0. 1 5 5 3 113:1:22 4 38. 011:5 1 32. 0=i=12 Non-parabiont female. 2 =P 0.025. 3 =P 0.05. P 0.005.

EXAMPLE 5 Preparation of tablets containing 2-acetylamino-5- nitrothiazole The anti-testosterone compound, Z-acetylamino-S-nitrothlazole is incorporated into a standard pharmaceutical tablet according to the following formulation.

For 10,000

Ingredient Per tablet, mg. tablets, g.

2-acetylamino-5-nitrothiazole 50 500 Lactose 225 2, 250 Corn starch (for mix) 50 500 Corn starch (for paste) 25 250 Magnesium stearate 50 500 Total 400 4,000

The active ingredient, Z-acetylamino-S-nitrothiazole, lactose and corn starch for mix are blended together. The corn starch for paste is suspended in water at a ratio of g. of corn starch per 800 ml. of water and heated with stirring to form a paste. The resulting paste is then used to granulate the blended powder. Additional water is used if necessary. The wet granules are passed through a No. 8 screen and dried at F. The dry granules are passed through a No. 16 screen and lubricated with the magnesium stearate. The granulation is then compressed into tablets (tablet wt. 355 mg.) in a suitable tableting machine. Each tablet contains 50 mg. of active ingredient.

EXAMPLE 6 Preparation of hard shell capsules containing 2-acetylamino-S-nitrothiazole The active ingredient, 2-acetylamino-S-nitrothiazole may be dispensed in hard shell capsules. A formulation found useful for preparing such capsules is as follows:

Per capsule, For 1,000

Ingredient mg. capsules, g.

Z-acetyiamino-fi-nltrothiazole 50 50 Lactose 90 90 Magnesium stearate. 1 1

The active ingredient, lactose and magnesium stearate are blended together. The mixture is used to fill hard shell capsules of a suitable size at 'a fill weight of 141 mg. per

capsule.

EXAMPLE 7 Preparation of oral suspension containing 2-propionylamino-S-nitrothiazole Ingredient: Amount, percent w./v. 2-propionylamino-S-nitrothiazole 1.00 Magnesium aluminum silicate gel 0.50 Sucrose 60.00 Methylparaben 0.08 Propylparaben 0.02

Flavoring agent, q.s. Distilled water, q.s. ad 100.00.

7 EXAMPLE 8 The active component, Z-acetylamino-S-nitrothiazole, may be incorporated into an injectable suspension according to the following formulation:

Ingredient: Amount, percent w./v. 2-acetylamino-S-nitrothiazole (micronized) 5.0 Polyethylene glycol 4000 U.S.P. 4.0 Sodium chloride U.S.P. 0.90 Benzyl alcohol, reagent grade 0.90

Water for injection, q.s. ad 100.0.

EXAMPLE 9 The anti-testosterone compound Z-acetylamino-S-nitm thiazole may be incorporated into a topical jelly according to the following formulation.

Ingredient: Amount, percent w./v. Z-acetylamino-S-nitrothiazole 1.0 Methocel 4000 2.75 Parabens (4:1 mixture of methyl and propyl) 0.1 Propylene glycol 20.0

Distilled water, q.s. 100.0.

The parabens and Methocel are dissolved in about twothirds of the final volume of distilled water at 80 C. with stirring. The gel is cooled to below 40 C. and is then added with stirring to a solution of the active ingredient in propylene glycol. The final gel is adjusted to volume with distilled water.

A ml. portion of this gel contains 50 mg. of 2-acetylamino-S-nitrothiazole.

EXAMPLE 10 A cream incorporating 2-acetylamino-S-nitrothiazole may be formulated as follows:

Ingredient: Amount, percent w./v.

2 acetylamino S-nitrothiazole (finely divided) 20.0

Liquid petrolatum 20.0

Cetyl alcohol 5.0 Parabens (4: 1 mixture of methyl and propyl) 0.1 Emulsifier 5.0 Glycerin 5.0 Distilled water, q.s. 100.0.

The parabens are dissolved in about two-thirds of the final volume of distilled water at C. with stirring. The solution is cooled to about 50 C. and glycerin is added. Light liquid petrolatum, cetyl alcohol and emulsifier are heated together to about 50 C., blended well, and added with stirring to the warm aqueous solution. The emulsion is cooled to room temperature and the active ingredient is then added with stirring. The volume is adjusted with distilled water.

I claim:

1. A method of decreasing sebum output in pubescent human beings which comprises topically administering to said pubescent human beings an effective sebum reducing amount of a nitrothiazole of the formula:

wherein R is loweralkanoyl.

2. A method in accordance with claim 1, in which the sebum reducing effect is produced by topically applying a composition containing as the essential active component 2-acetylamino-S-nitrothiazole.

3. A method in accordance with claim 2, wherein the composition contains at least 25% of 2- acetylamino-5- nitrothiazole.

4. A method of decreasing sebum output in human beings which comprises topically applying to the intact skin of said human beings a composition containing from about 1 mg. to 64 mg. per kilogram of human being of 2-acetylamino-5-nitrothiazole.

5. A topical composition for decreasing the sebum output in pubescent human beings which comprises a pharmaceutical carrier and from 25 mg. to 250 mg. of 2-lower alkanoylamino-S-nitrotlfiazole.

6. A topical composition in accordance with claim 5, in which the active component is 2 acetylamino 5- nitrothiazole.

References Cited UNITED STATES PATENTS 2,531,756 11/1950 Waletsky et al 424263 OTHER REFERENCES Chem. A'bsL, 58, 1448l (b), 1963.

STANLEY I FRIEDMAN, Primary Examiner 

